• 00:00 1.
    Comparative Gene-expression Analysis
  • 00:15 2.
    Question
  • 00:40 3.
    Differential Display
  • 01:20 4.
    Polymerase Chain Reaction
  • 04:15 5.
    Template DNA
  • 07:24 6.
    SpecificityMust be complementary to flanking sequences of target regionNot be complementary to other non-target regions of genomeNested polymerase chain reaction (nested PCR): to reduce non-specific binding Primer LengthToo short  lack specificityToo lon
  • 08:56 7.
    Product SizeBasic Taq polymerase can easily amplify fragments up to 1,000 to 2,000 bpTypical qPCR amplicons are 70-200 bp in lengthToo long  Fluorescent intensity not in linear rangePrimer DimersIf the primers have self-complementary sequences, the prime
  • 11:35 8.
    General Considerations in Primer Design
  • 16:45 9.
    General Considerations in Primer Design
  • 18:02 10.
    G/C ContentPrimers should be about 50% G/C (40~60%) Not have long runs of G/C or A/T (>3 bp) A stretch of A/T’s might only weakly base pairA stretch of G/C might promote mis-annealingG/C clampTo ensure the stability of this interaction, primers are often
  • 18:45 11.
    Web Based Tools for Primer Design
  • 18:52 12.
    http://biotools.nubic.northwestern.edu/OligoCalc.html
  • 19:02 13.
    Web Based Tools for Primer Design
  • 19:08 14.
    https://www.ncbi.nlm.nih.gov/tools/primer-blast/
  • 19:13 15.
    Question
  • 20:23 16.
    Rapid Amplification of cDNA Ends (RACE)
  • 21:50 17.
    Rapid Amplification of cDNA Ends (RACE)
  • 22:51 18.
    Multiplex PCR
  • 25:04 19.
    Primer Design Assistant
  • 25:13 20.
    PCR-based Techniques: Subtractive hybridization
  • 28:41 21.
    QuestionCan RNA be amplified directly by PCR? Why and why not?
  • 29:39 22.
    Reverse Transcription PCR
  • 31:40 23.
    Real Time - PCR
  • 34:45 24.
    Real Time - PCR
  • 37:27 25.
    Amplification Plot
  • Index
  • Notes
  • Comment
  • Fullscreen
Comparative Gene-Expression Analysis II: PCR-based techniques (0323)
Duration: 41:55, Browse: 519, Last Updated: 2021-03-30
    • 00:00 1.
      Comparative Gene-expression Analysis
    • 00:15 2.
      Question
    • 00:40 3.
      Differential Display
    • 01:20 4.
      Polymerase Chain Reaction
    • 04:15 5.
      Template DNA
    • 07:24 6.
      SpecificityMust be complementary to flanking sequences of target regionNot be complementary to other non-target regions of genomeNested polymerase chain reaction (nested PCR): to reduce non-specific binding Primer LengthToo short  lack specificityToo lon
    • 08:56 7.
      Product SizeBasic Taq polymerase can easily amplify fragments up to 1,000 to 2,000 bpTypical qPCR amplicons are 70-200 bp in lengthToo long  Fluorescent intensity not in linear rangePrimer DimersIf the primers have self-complementary sequences, the prime
    • 11:35 8.
      General Considerations in Primer Design
    • 16:45 9.
      General Considerations in Primer Design
    • 18:02 10.
      G/C ContentPrimers should be about 50% G/C (40~60%) Not have long runs of G/C or A/T (>3 bp) A stretch of A/T’s might only weakly base pairA stretch of G/C might promote mis-annealingG/C clampTo ensure the stability of this interaction, primers are often
    • 18:45 11.
      Web Based Tools for Primer Design
    • 18:52 12.
      http://biotools.nubic.northwestern.edu/OligoCalc.html
    • 19:02 13.
      Web Based Tools for Primer Design
    • 19:08 14.
      https://www.ncbi.nlm.nih.gov/tools/primer-blast/
    • 19:13 15.
      Question
    • 20:23 16.
      Rapid Amplification of cDNA Ends (RACE)
    • 21:50 17.
      Rapid Amplification of cDNA Ends (RACE)
    • 22:51 18.
      Multiplex PCR
    • 25:04 19.
      Primer Design Assistant
    • 25:13 20.
      PCR-based Techniques: Subtractive hybridization
    • 28:41 21.
      QuestionCan RNA be amplified directly by PCR? Why and why not?
    • 29:39 22.
      Reverse Transcription PCR
    • 31:40 23.
      Real Time - PCR
    • 34:45 24.
      Real Time - PCR
    • 37:27 25.
      Amplification Plot
    Location
    Folder name
    2021
    Author
    賴亮全
    Branch
    賴亮全教授
    Created
    2021-03-23 22:40:17
    Last Updated
    2021-03-30 20:16:27
    Duration
    41:55